The prior art is replete with a variety of compositions directed toward the effective killing of bacterial spores. Among the most important developments in this area are the compositions disclosed in U.S. Pat. No. 3,016,328 to Pepper et al and U.S. Pat. No. 3,282,775 to Stonehill. In the former patent saturated dialdehydes provide sporicidal activity and the pH of the composition is controlled so that it is maintained in excess of 7.4 by including an alkalinizing agent. The sporicidal composition disclosed in the latter patent to Stonehill is also characterized as containing a saturated dialdehyde, including cationic surface active agents. In the Stonehill patent, it is expressly stated that anionic and/or nonionic detergents do not increase the sporicidal activity of the compositions.
While the patent to Pepper et al limits the pH of the sporicidal composition to a minimum in excess of at least 7.4, the Stonehill patent discloses no such limitation as the compositions disclosed therein are stated to be effective over a wide pH range of 4.0-9.0.
Each of the sporicidal compositions disclosed in the above-identified patents claim very fast sporicidal action of about 3 hours or less. However, close examination of the conditions upon which such fast kill claims were determined reveals that the "kills" were obtained against readily susceptible bacterial spores and that neither patent discloses any sporicidal activity results for bacterial spores on silk sutures, which carrier is specified as being required by the AOAC test procedure. Furthermore, it is well known that resistance of bacterial spores to chemical sterilizing agents is lowest in free suspension, intermediate on porcelain rings and most resistant on silk sutures. Recently, the claims for sporicidal compositions marketed by the common assignee of the above-noted patents had to be revised in their registration with the USDA from a contact kill time of 3 hours upward to 10 hours.
Independent analyses of the sporicidal compositions disclosed in U.S. Pat. No. 3,016,328 to Pepper et al revealed that the 10 hour contact kill time was readily obtainable when using a fresh solution, but that the efficacy of the compositions markedly decreased upon standing for prolonged periods of up to about 2 weeks. Further, this reduction in effectiveness was found to be attributable to the diminution of glutaraldehyde, which lost a total of about 25% of its value by the end of a two week period.
It can be seen, therefore, that these two principal disclosures relating to sporicidal compositions, under the limited test conditions set forth therein may not, in reality, exhibit the effectiveness implied for them with respect to passing the complete AOAC test. Furthermore, the effectiveness of the disclosed compositions are based upon those bacterial spores and/or carriers which are known to favor relatively easy "kills."
It has now been found that the shortcomings of the prior art can be overcome by use of the glutaraldehyde sporicidal compositions of the invention which employ the combination of glutaraldehyde and monoaldehyde, such as formaldehyde. The compositions provide a minimum amount of glutaraldehyde in a suitable sporicidal solvent and may further include an anionic, nonionic, cationic or ampholytic detergent therein to obtain enhanced sporicidal activity. Further, by closely controlling the pH of the compositions, significantly improved shelf life is provided which is manifested by the sporicidal performance of the activated compositions, even after standing for prolonged periods of 2 to 3 weeks.
The amount of glutaraldehyde incorporated in the sporicidal solvent should be no less than about 0.5% by weight, since lesser amounts unduly prolong the kill times, while the maximum amount which can be used is essentially without limit. The term "sporicidal solvent," as used throughout this application and in the claims, should be understood as referring to those solvents normally employed for sporicidal compositions and which include water and/or alcohols. For example, the U.S. Patents to Pepper et al and Stonehill et al discussed above, each disclose the use of alcohols as a sporicidal solvent. However, in this invention, water is the preferred sporicidal solvent to be used, although other sporicidal solvents can also be employed.
The addition of activating agents to adjust the pH of sporicidal compositions is well known to those skilled in the art. Generally, glutaraldehyde compositions are stable almost indefinitely within the pH range of about 2.5-4.5 at which pH levels they are stored before use. Just prior to use, their pH levels are adjusted through the addition of activating agents. Hence, most sporicidal compositions are made commercially available as a two package system, one of which comprises the sporicide in a suitable solvent and the other of which contains the activating agent, either as a powder or in solution, which is to be added to the sporicidal composition to activate it and adjust its pH just prior to use. The addition of such agents has been noted above in discussing the patent to Pepper et al (U.S. Pat. No. 3,016,328) wherein this procedure is referred to as "alkalinizing" the composition. In the context of this invention, the term "activating" is employed and should be understood to be equivalent to alkalinizing such compositions and adjusting their pH levels by adding well known buffering agents to them. Once activated in this manner, the sporicidal compositions of the prior art have been found to have limited shelf lives and are, therefore, generally utilized immediately, or within a very short time, after being activated. It is to this particular problem that one embodiment of this invention is directed for it has now been found that upon activating the sporicidal compositions of this invention so that their pH levels are maintained within a specified range, improved shelf life stability can be obtained.
The pH of the activated sporicidal compositions of the invention can be controlled by incorporating therein one or more of the suitable and well known buffering agents so that the pH of the composition is no greater than 7.4, preferably about 6.5 to 7.4, and optimumly at a pH of 7.0 .+-. 0.3. The selection of suitable buffering agents for controlling the pH level is not critical and such materials as phosphates, citrates, carbonates, bicarbonates and the like, can be readily employed, although the phosphates are particularly preferred due to their favorable dissociation constants. As is well known in the art, other ingredients such as anti-corrosion agents, dyes, and the like, can also be added to the compositions.
In the principal embodiment of the invention it has been found that the inclusion of one or more monoaldehydes results in a synergistic effect of greatly reduced kill times thereby further enhancing the efficacy of the composition. Such compositions are especially effective for killing Bacillus subtilis (B. subtilis), one of the acknowledged most difficult bacterial spores to kill. The monoaldehydes should be present in amounts no less than about 0.5% with the upper amounts being limited only by their solubility in the sporicidal solvent being employed. Illustrative of the monoaldehydes which can be employed are formaldehyde, acetaldehyde, propionaldehyde, and butyraldehyde, formaldehyde being preferred.
The detergents which can additionally be employed in the composition are not limited to any one of the known detergent classes or groups, and it has been found that their use serves to potentiate; that is, increase and enhance, the sporicidal activity of the compositions. The minimum amount of detergent which should be employed is about 0.01% by weight with a range of about 0.1 to 1.0% preferred. For the purposes of this invention the term "detergent" should be understood as referring to any substance which, when added to water at a concentration of 0.1%, will depress the surface tension of water by at least 20 dynes per square centimeter. Under this criterion, any nonionic, anionic, cationic, and/or ampholytic detergents can be utilized. When exposed to some materials, such as metal instruments; for example, scalpels, anionic detergents may exhibit a corrosive effect, and for this reason the nonionic detergents are preferred. Exemplary of the nonionic detergents which can be employed are the alkylphenolethoxylates available uner the Trademark "Igepal."
The sporicidal compositions of the invention have been found to be effective in killing a wide range of bacterial spores such as Clostridium welchii (Cl. welchii), Clostridium tetani (Cl. tetani), Bacillus subtilis (B. subtilis), Bacillus pumilus (B. pumilus), Bacillus globigii (B. globigii), Clostridium sporogenes (Cl. sporogenes), and the like. Of these, the Cl. sporogenes and the B. subtilis are known to be among the most difficult bacterial spores to kill, and are the organisms specified in the AOAC test.